Maternal obesity alters brain derived neurotrophic factor (BDNF) signaling in the placenta in a sexually dimorphic manner  Calais S. Prince, Alina Maloyan, Leslie Myatt  Placenta  Volume 49, Pages 55-63 (January 2017) DOI: 10.1016/j.placenta.2016.11.010 Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 1 miR-210 and BDNF mRNA expression in placentas from lean and obese women. Quantification of miR-210 (A), BDNF Exon IVS (B), and Exon VS (D) by RT-PCR. U18 was used as an internal control for miR-210 and ribosomal 18S was used as an internal control for BDNF mRNA. Data were fitted using linear regression analysis. Correlation coefficient (R) and p values are shown. Correlation analysis between miR-210 expression, BDNF Exon IVS expression (C), and BDNF Exon VS expression (E) in placentas from lean and obese women carrying either a male or a female fetus. Values from miR-210 RT-PCR are mean ± SEM; a: p = 0.01 vs. LN female; n = 13 per group/sex. Values from BDNF RT-PCR and mean ± SEM; *: p = 0.0018 vs LN; a: p = 0.012 vs LN female; n = 13 per group/sex. Dotted regression line: no correlation (p > 0.05). Placenta 2017 49, 55-63DOI: (10.1016/j.placenta.2016.11.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 2 ProBDNF levels and correlation with miR-210 expression in placentas from lean and obese women. ProBDNF protein in placental homogenates from lean or obese women carrying either a male or female fetus (A). ProBDNF was normalized to total protein. Data were fitted using linear regression analysis. Correlation coefficient (R) and p values are shown. Correlation between miR-210 expression and proBDNF levels in placentas from lean and obese women with male or female fetuses (B), placentas from lean and obese women carrying a male fetus (C), and placentas from lean and obese women carrying a female fetus (D). Values from the BDNF ELISA are median with minimum and maximum values, *: p = 0.02 vs LN; a: p = 0.03 vs. LN male; n = 10 per group/sex. Dotted regression line: no correlation (p > 0.05). Placenta 2017 49, 55-63DOI: (10.1016/j.placenta.2016.11.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 3 BDNF protein levels and correlation with miR-210 in placentas from lean and obese women. Representative Western blot (A) and quantification (B) of mature BDNF in placentas from lean and obese women carrying either a male or female fetus. β-Actin was used as the loading control. Data were fitted using linear regression analysis. Correlation coefficient (R) and p values are shown. Correlation between miR-210 expression and BDNF levels in placentas from lean and obese women carrying either a male or female fetus (C). Correlation between miR-210 expression and BDNF levels in placentas from lean women carrying either a male or female fetus (D). Correlation between miR-210 expression and BDNF levels in placentas from obese women carrying either a male or female fetus (E). Values from Western blot are mean ± SEM; *:p = 0.02 vs female; a: p = 0.04 vs LN female; n = 10 per group/sex. Dotted regression line: no correlation (p > 0.05); solid regression line: correlation (p = 0.04). Placenta 2017 49, 55-63DOI: (10.1016/j.placenta.2016.11.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 4 TRKB levels and TRKB phosphorylation at tyrosine 515 and 817 in placentas from lean and obese women with male or female fetuses. Representative Western blots (A, D) and quantification of pY515 TRKB (B, C), pY817 TRKB (E, F), and TRKB (G). β actin was used as the loading control. Values are mean ± SEM; *: p < 0.05 vs. LN; n = 10 per group/sex. Placenta 2017 49, 55-63DOI: (10.1016/j.placenta.2016.11.010) Copyright © 2016 Elsevier Ltd Terms and Conditions

Fig. 5 p38 MAPK and p42/44 MAPK phosphorylation in placentas from lean and obese women with male or female fetuses. Representative Western blots and quantification of pp38 MAPK (A, B) and pp42/44 MAPK (C, D). Samples were normalized to p38 and p42/44 respectively. β actin was used as the loading control. Values are mean ± SEM; *: p = 0.01 vs. LN; a: p = 0.01 vs LN male; n = 6 per group/sex (pp38); n = 13 per group/sex (pp42/44). Placenta 2017 49, 55-63DOI: (10.1016/j.placenta.2016.11.010) Copyright © 2016 Elsevier Ltd Terms and Conditions